Digestion of whey peptide induces antioxidant and anti-inflammatory bioactivity on glial cells: Sequences identification and structural activity analysis.

Whey derived peptides have shown potential activity improving brain function in pathological condition. However, there is little information about their mechanism of action on glial cells, which have important immune functions in brain. Astrocytes and microglia are essential in inflammatory and oxidative defense that take place in neurodegenerative disease. In this work we evaluate antioxidant and anti-inflammatory potential bioactivity of whey peptide in glial cells. Peptides were formed during simulated gastrointestinal digestion (Infogest protocol), and low molecular weight (<5kDA) peptides (WPHf) attenuated reactive oxygen species (ROS) production induced by hydrogen peroxide stimulus in both cells in dose-dependent manner. WPHf induced an increase in the antioxidant glutathione (GSH) content and prevented GSH reduction induced by lipopolysaccharides (LPS) stimulus in astrocytes cells in a cell specific form. An increase in cytokine mRNA expression (TNFα and IL6) and nitric oxide secretion induced by LPS was attenuated by WPHf pre-treatment in both cells. The inflammatory pathway was dependent on NFκB activation. Bioactive peptide ranking analysis showed positive correlation with hydrophobicity and negative correlation with high molecular weights. The sequence identification revealed 19 peptides cross-referred with bioactive database. Whey peptides were rich in leucine, valine and tyrosine in the C-terminal region and lysine in the N-terminal region. The anti-inflammatory and antioxidant potential of whey peptides were assessed in glia cells and its mechanisms of action were related, such as modulation of antioxidant enzymes and anti-inflammatory pathways. Features of the peptide structure, such as molecular size, hydrophobicity and types of amino acids present in the terminal region are associated to bioactivity.

Formulations with microencapsulated Fe-peptides improve in vitro bioaccessibility and bioavailability.

The bioaccessibility and the bioavailability of iron complexed to peptides (active) in microparticles forms contained in dry beverages formulations were evaluated. The peptide-iron complexes microparticles were obtained by spray drying and added in three dry formulations (tangerine, strawberry, and chocolate flavors). The peptides isolated by iron ion affinity (IMAC-Fe III) had their biological activity predicted by BIOPEP® database and were evaluated by molecular coupling. The bioaccessibility was evaluated by solubility and dialysability and the bioavalability was assessed by Caco-2 cellular model. The proportion 10:1 of peptide-iron complexes presented higher rates of bioaccessibility (49%) and bioavailability (56%). The microparticle with peptide-iron complex showed greater solubility after digestion (39.1%), bioaccessibility (19.8%), and bioavailability (34.8%) than the ferrous sulfate salt (control) for the three assays (10.2%; 12.9%; 9.7%, respectively). Tangerine and strawberry formulations contributed to the iron absorption according to the results of bioaccessibility (36.2%, 30.0% respectively) and bioavailability (80.5%, 84.1%, respectively). The results showed that iron peptide complexation and microencapsulation process improve the bioaccessibility and bioavailability when incorporated into formulations.

Amorphophallus konjac: A Novel Alternative Flour on Gluten-Free Bread.

The demand for gluten-free products is rising, but their production with similar quality as their gluten counterparts is challenging. This study aimed to develop gluten-free bread samples using different concentrations of Amorphophallus konjac flour (0%, 12.5%, 25%, 37.5%, and 50% of the total flour content) and to evaluate their nutritional and physicochemical properties. Proteins, lipids, carbohydrates, moisture, ash content, fibers, resistant starch, firmness, specific volume, and color were evaluated using official methods. Protein varied from 2.95% to 4.94%, the energy value from 347.93 to 133.55 kcal/100 g, dietary fiber from 8.19 to 17.90%, and resistant starch from 0.67% to 0.75% on wet basis. The addition of konjac flour positively influenced the specific volume. Higher concentrations of konjac flour in the formulations led to lower calories of the bread due to the significant addition of water to the dough. The bread samples with konjac showed high fiber content due to the composition of the flour. They had lower levels of carbohydrates, which can positively influence the glycemic index. Konjac flour provided dough mold, growth, and better texture for gluten-free bread. The best formulations were prepared in concentrations up to 37.5% konjac. The 50% konjac bread showed slightly reduced specific volume and pale color.

Identification of bioactive peptides released from in vitro gastrointestinal digestion of yam proteins (Dioscorea cayennensis).

Bioactive peptides have been broadly studied for their contribution to human health. This study aimed to identify bioactive peptides generated by in vitro gastrointestinal digestion of yam proteins. Yam protein concentrate (YPC) was submitted to simulated digestion. Gastric phase hydrolysate (GPH) and total gastrointestinal phase hydrolysate (GIPH) had their peptides identified by nanoLC-ESI-MS/MS. Peptide sequences were subjected to a database-driven (BIOPEP) bioactivity search. In vitro tests included: Antioxidant activity, DNA damage protection, ACE-inhibitory activity and antibacterial activity against the bacteria Escherichia coli, Salmonella sp. and Lysteria monocytogenes. Simulated digestion generated small peptides (mostly MW < 3500 Da), several of them with potential bioactive sequences predicted in silico. In both GPH and GIPH biological activities were detected, although GIPH displayed stronger DNA damage protection and antibacterial activity against Escherichia coli. The digestion of yam proteins releases promising biologically active peptides which can contribute to the prevention of bacterial infection and chronic degenerative diseases, with beneficial effects to human health.

Bioaccessibility of cashew nut kernel flour compounds released after simulated in vitro human gastrointestinal digestion.

Cashew nuts are mainly consumed as a roasted and salted snack. Lately, the industry has gained interest in broken kernels because of their added value. In this study, defatted cashew nut flour (DCF) underwent simulated gastrointestinal digestion to obtain a soluble (CDs) and an insoluble (CDi) digested fraction. These fractions, which resulted from the digestion of a complex matrix, were evaluated for antioxidant capacity of bioaccessible compounds (present on the soluble digested fraction, CDs) and their potential prebiotic effect, considering that the insoluble digested fraction (CDi) could be fermented by the microbiota in the gut. The DCF had a high protein content (40.74%), being nutritionally characterized as a balanced source of amino acids, with a predominance of aromatic amino acids (phenylalanine and tyrosine), threonine and histidine. The digested DCF presented 76.90% of the soluble components of low molecular weight (0.1-2 kDa), which is typical of antioxidant peptides. The soluble digested fraction (CDs) significantly increased the antioxidant capacity in relation to flour in the ORAC and ABTS assays and the aqueous extract presented the highest values (526.0 and 76.64 as µmol Trolox Eq./g sample, respectively). The CDs protected 29.03% of the supercoiled DNA band and ratified the potential antioxidant capacity after GID in a physiological assay. In addition, the insoluble digested fraction showed a potential prebiotic effect for Bifdobacterium lactis BB-12. Finally, simulated gastrointestinal digestion improves the bioaccessibility of CDF antioxidant compounds as a complex matrix, containing low molecular weight peptides and phenolic compounds, which become more available to react with reactive oxygen species (ROS). In addition, the potential prebiotic effect of defatted cashew nut flour has yielded a promising solution for the total reuse of broken cashew nut kernel as a functional food ingredient.

Peptide-metal complexes: obtention and role in increasing bioavailability and decreasing the pro-oxidant effect of minerals.

Bioactive peptides derived from food protein sources have been widely studied in the last years, and scientific researchers have been proving their role in human health, beyond their nutritional value. Several bioactivities have been attributed to these peptides, such as immunomodulatory, antimicrobial, antioxidant, antihypertensive, and opioid. Among them, metal-binding capacity has gained prominence. Mineral chelating peptides have shown potential to be applied in food products so as to decrease mineral deficiencies since peptide-metal complexes could enhance their bioavailability. Furthermore, many studies have been investigating their potential to decrease the Fe pro-oxidant effect by forming a stable structure with the metal and avoiding its interaction with other food constituents. These complexes can be formed during gastrointestinal digestion or can be synthesized prior to intake, with the aim to protect the mineral through the gastrointestinal tract. This review addresses: (i) the amino acid residues for metal-binding peptides and their main protein sources, (ii) peptide-metal complexation prior to or during gastrointestinal digestion, (iii) the function of metal (especially Fe, Ca, and Zn)-binding peptides on the metal bioavailability and (iv) their reactivity and possible pro-oxidant and side effects.

Whey Peptide-Iron Complexes Increase the Oxidative Stability of Oil-in-Water Emulsions in Comparison to Iron Salts.

Food fortification with iron may favor lipid oxidation in both food matrices and the human body. This study aimed at evaluating the effect of peptide-iron complexation on lipid oxidation catalyzed by iron, using oil-in-water (O/W) emulsions as a model system. The extent of lipid oxidation of emulsions containing iron salts (FeSO4 or FeCl2) or iron complexes (peptide-iron complexes or ferrous bisglycinate) was evaluated during 7 days, measured as primary (peroxide value) and secondary products (TBARS and volatile compounds). Both salts catalyzed lipid oxidation, leading to peroxide values 2.6- to 4.6-fold higher than the values found for the peptide-iron complexes. The addition of the peptide-iron complexes resulted in the formation of lower amounts of secondary volatiles of lipid oxidation (up to 78-fold) than those of iron salts, possibly due to the antioxidant activity of the peptides and their capacity to keep iron apart from the lipid phase, since the iron atom is coordinated and takes part in a stable structure. The peptide-iron complexes showed potential to reduce the undesirable sensory changes in food products and to decrease the side effects related to free iron and the lipid damage of cell membranes in the organism, due to the lower reactivity of iron in the complexed form.

Synthesis of whey peptide-iron complexes: Influence of using different iron precursor compounds.

Iron-binding peptides are an alternative for increasing the bioavailability of iron and to decreasing its pro-oxidant effect. This study aimed to synthesize and characterize peptide-iron complexes using FeCl2 or FeSO4 as the iron precursor compounds. Whey protein isolate (WPI), WPI hydrolyzed with pancreatin, and its fractions obtained via ultrafiltration (cut-off 5kDa) were used as ligands. The fluorescence intensity of the ligands significantly decreased as the iron concentration increased as a result of metal coordination with the iron-binding sites, which may have led to changes in the microenvironment of tryptophan. For both iron precursor compounds, the primary iron-binding site was carboxylate groups, and the linkage occurred via a bidentate coordination mode with two vibrational modes assigned to the COOFe linkage. However, infrared spectroscopy and thermal analysis results showed that the dynamics of the interaction is different for the iron precursor. The iron source may be of great importance because it may impact iron absorption and the pro-oxidant effect of the mineral.

Iron-binding properties of sugar cane yeast peptides.

The extract of sugar-cane yeast (Saccharomyces cerevisiae) was enzymatically hydrolysed by Alcalase, Protex or Viscozyme. Hydrolysates were fractionated using a membrane ultrafiltration system and peptides smaller than 5kDa were evaluated for iron chelating ability through measurements of iron solubility, binding capacity and dialyzability. Iron-chelating peptides were isolated using immobilized metal affinity chromatography (IMAC). They showed higher content of His, Lys, and Arg than the original hydrolysates. In spite of poor iron solubility, hydrolysates of Viscozyme provided higher iron dialyzability than those of other enzymes. This means that more chelates of iron or complexes were formed and these kept the iron stable during simulated gastro-intestinal digestion in vitro, improving its dialyzability.